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Immunology and Infection
İnsan THP-1 Makrofajlarında Candida glabrata'nın Hücre İçi Davranışını İncelemek İçin
İnsan THP-1 Makrofajlarında Candida glabrata'nın Hücre İçi Davranışını İncelemek İçin
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Establishment of an In vitro System to Study Intracellular Behavior of Candida glabrata in Human THP-1 Macrophages

İnsan THP-1 Makrofajlarında Candida glabrata'nın Hücre İçi Davranışını İncelemek İçin İn vitro Sistemin Kurulması

Full Text
12,266 Views
13:47 min
December 10, 2013

DOI: 10.3791/50625-v

Maruti Nandan Rai1, Sapan Borah1, Gaurav Bairwa*1, Sriram Balusu*1,2, Neelima Gorityala1, Rupinder Kaur1

1Laboratory of Fungal Pathogenesis,Centre for DNA Fingerprinting and Diagnostics, Andhra Pradesh, India, 2Current location: VIB Department for Molecular Biomedical Research, UGent,Fiers-Schell-Van Montagu Building, Technologiepark 927, B-9052 Ghent (Zwijnaarde), Belgium

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Overview

This article presents a protocol for establishing an in vitro cell culture model to investigate the interaction between the human fungal pathogen Candida glabrata and human macrophages. This model aims to enhance understanding of fungal virulence mechanisms.

Key Study Components

Area of Science

  • Microbiology
  • Immunology
  • Cell Biology

Background

  • Candida glabrata is a facultative intracellular pathogen.
  • Understanding its interaction with macrophages is crucial for insights into fungal infections.
  • THP-1 cell line is commonly used for macrophage differentiation.
  • In vitro models are essential for studying pathogen-host interactions.

Purpose of Study

  • To develop a reliable in vitro model for studying Candida glabrata.
  • To assess the intracellular behavior of the pathogen in human macrophages.
  • To evaluate macrophage responses to Candida infection.

Methods Used

  • THP-1 monocytes are differentiated into macrophages using PMA treatment.
  • Macrophages are infected with Candida glabrata cells.
  • Non-phagocytosed cells are removed through PBS washes.
  • Intracellular cells are recovered for further analysis.

Main Results

  • The protocol allows for the assessment of Candida glabrata virulence in macrophages.
  • It enables the study of cytokine responses in THP-1 macrophages.
  • Insights into the maturation processes of macrophages upon infection are provided.

Conclusions

  • This in vitro model is a valuable tool for studying fungal pathogenesis.
  • It contributes to the understanding of host-pathogen interactions.
  • Future studies can leverage this model to explore therapeutic interventions.

Frequently Asked Questions

What is the significance of studying Candida glabrata?
Candida glabrata is an opportunistic pathogen that poses a significant threat to immunocompromised individuals, making its study crucial for developing effective treatments.
How are THP-1 cells differentiated into macrophages?
THP-1 cells are treated with PMA to induce differentiation into macrophage-like cells.
What techniques are used to analyze intracellular Candida glabrata?
The protocol includes washing non-phagocytosed cells and recovering intracellular cells for further analysis.
What are the expected outcomes of this study?
The study aims to elucidate the virulence mechanisms of Candida glabrata and the immune response of macrophages.
Can this model be used for other pathogens?
Yes, the model can potentially be adapted to study other intracellular pathogens.
What is the role of cytokines in this study?
Cytokines are key indicators of the immune response and are measured to assess macrophage activation during infection.

Bu makale, mantar virülans mekanizmaları hakkındaki bilgimizi ilerletmek için yararlı bir araç olacak olan fakültatif bir hücre içi insan mantar patojeni Candida glabrata'nın insan makrofajları ile etkileşimini incelemek için bir in vitro hücre kültürü model sistemi oluşturmak için bir protokolün ana hatlarını çizmektedir.

Bu prosedürün genel amacı, bir insan fırsatçı mantar patojeninin hücre içi davranışını incelemek için bir in vitro kültür modeli sistemi oluşturmaktır. İnsan monositik hücre hattı TP'den türetilen makrofajlarda Canida lab ratta. Bu THP'yi elde etmek için, bir monosit önce onları makrofajlara ayırmak için dört ester ile muamele edilir.

Daha sonra, bir makrofaj, enfeksiyondan iki saat sonra canida GTA hücreleri ile enfekte olur. Fagositoz olmayan canida GTA hücreleri PBS yıkayıcıları ile uzaklaştırılır ve hücre içi e hücreleri, enfekte makrofajların ozmozu ile daha ileri çalışmalar için geri kazanılır. Bu protokol, hem makrofajlardaki viral to-canida GTA hücrelerini hem de candida GTA enfeksiyonu üzerine makro FIDS'de faiz maturasyonu Ross ve THP'deki sitokin yanıtını değerlendirmek için kullanılabilir.

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Anahtar Kelimeler: İn Vitro Model THP-1 Makrofajları Candida Glabrata Hücre İçi Patojen Konak-Patojen Etkileşimi Mikrobiyal Virülans Hücre Kültürü Enfeksiyon Modeli Yüksek Verimli Tarama

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